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May 15, 2018 at 14:42 #1773
Dear all, together with my student Hadar Amartely we develop a new method for protein characterization. The method was published last week in Scientific Reports: Coupling Multi Angle Light Scattering to Ion Exchange chromatography (IEX-MALS) for protein characterization.
The article is freely available online at http://www.nature.com/articles/s41598-018-25246-6
IEX-MALS is a valuable and complementary method for protein characterization for Academy and Pharma, especially for protein systems that could not be fully analyzed by SEC-MALS.
We show examples of oligomers that cannot be resolved by SEC-MALS, and we succeed to do it by IEX-MALS (example of: dimer-tetramer-hexamer-octamer). Moreover, aggregates elute after the monomer, minimizing contamination of the monomer signal. We succeed to analyze the oligomeric conformation of peptides. Single peaks in SEC-MALS can be resolve in multiple peaks in IEX-MALS (examples of therapeutic proteins), etc
The limited resolution of SEC restricts in some instances the accurate analysis that can be accomplished by MALS. IEX-MALS allows more precise analyses of some samples that cannot be resolved by SEC-MALS. Main advantages:
• Different principle of separation (Analysis of mixtures of proteins with similar size) Therefore it can be used for analysis of mix proteins samples (not only pure samples)
• Huge variety of parameters that increase selectivity and resolution: Different steps/gradient running programs, gradient slope, pH or salt gradient, type of salts, type of bufer, resin particle size, diferent matrixes, type of column (CIEX/AIEX), column length and column supplier
• Elution of high order oligomers after monomers, dimers. This allows minimal contamination of the low MW signals
• Unlimited injection volume. Large quantities of peptides or small proteins can be loaded onto the column to improve signal-to-noise ratio and, ultimately, molar mass measurement—especially important when the sample is aggregation-prone and cannot be concentrated
• We conclude that IEX-MALS is a valuable and complementary method for protein characterization for Academy and Pharma
More information you can see on the blog of WYATT: https://bit.ly/2wIco1d
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